title: Characterization, expression and localization of valosin-containing protein in ovaries of the giant tiger shrimp Penaeus monodon.
abstract: Valosin-containing protein (VCP), a member of the ATPase-associated with diverse cellular activity (AAA) family, was identified from the giant tiger shrimp (Penaeus monodon). The full-length cDNA of the PmVCP mRNA consisted of 2,724 bp containing an ORF of 2,367 bp corresponding to a deduced polypeptide of 788 amino acids. The deduced PmVCP protein contained two putative Cdc48 domains (positions 17-103, E-value=2.00e-36 and 120-186, E-value=3.60e-11) and two putative AAA domains (positions 232-368, E-value=3.67e-24 and 505-644, E-value=3.73e-25). PmVCP mRNA expression in ovaries was greater than that in testes in both juveniles and broodstock. PmVCP was significantly up-regulated in stages II and IV ovaries in intact wild broodstock (P<0.05) but it was not differentially expressed during ovarian development in eyestalk-ablated broodstock (P>0.05). The expression level of PmVCP mRNA in ovaries of 14-month-old shrimp was not affected by progesterone injection (0.1μg/g body weight, P>0.05). In contrast, exogenous 5-HT administration (50μg/g body weight) resulted in an increase of PmVCP mRNA in ovaries of 18-month-old shrimp at 6 and 24h post-injection (hpi) (P<0.05). The rPmCdc48-VCP protein and its polyclonal antibody were successfully produced. Cellular localization revealed that PmVCP was localized in the ooplasm of previtellogenic oocytes. Subsequently, it was translocated into the germinal vesicle of vitellogenic oocytes. Interestingly, PmVCP was found in nucleo-cytoplasmic compartments, in the cytoskeletal architecture and in the plasma membrane of mature oocytes in both intact and eyestalk-ablated broodstock.
authors.affiliation: Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.
authors.forename: Witchulada
authors.initials: W
authors.lastname: Talakhun
authors.forename: Bavornlak
authors.initials: B
authors.lastname: Khamnamtong
authors.forename: Pachumporn
authors.initials: P
authors.lastname: Nounurai
authors.forename: Sirawut
authors.initials: S
authors.lastname: Klinbunga
authors.forename: Piamsak
authors.initials: P
authors.lastname: Menasveta
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers; D005260:Female; D010053:Ovary / Q000378:metabolism*; D033561:Penaeidae / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D000074405:Valosin Containing Protein
chemical_list: D018797:Cell Cycle Proteins; D017931:DNA Primers; D012333:RNA, Messenger; D000251:Adenosine Triphosphatases; D000074405:Valosin Containing Protein
keywords: 5-HT; 5-hydroxytryptamine or serotonin; AAA; ATPase-associated with diverse cellular activity; Cdc48; Cp; GSI; GVBD; Gene expression; Immunofluorescence; Immunohistochemistry; MPF; Ovarian development; Shrimp; VCP; Valosin-containing protein; cell division cycle protein 48; crossing points of real-time PCR analysis; germinal vesicle breakdown; gonadosomatic index; maturation promotion factor; nano-electrospay liquid chromatography ionization tandem mass spectrometry; nanoESI-LC–MS/MS; valosin-containing protein